{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,9,1]],"date-time":"2023-09-01T11:49:24Z","timestamp":1693568964807},"reference-count":20,"publisher":"Cambridge University Press (CUP)","issue":"2","license":[{"start":{"date-parts":[[2005,5,19]],"date-time":"2005-05-19T00:00:00Z","timestamp":1116460800000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/www.cambridge.org\/core\/terms"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Parasitology"],"published-print":{"date-parts":[[2005,8]]},"abstract":"<jats:p>The goal of this work is to report a novel assay that preserves antigenicity of extracts with high protease content without using enzymatic inhibitors. A great reduction of proteolytic activity in the insoluble chloroform\/methanol and heated amoebic fraction (IC[ratio ]MC) was obtained by this method, even in the presence of sodium dodecyl sulphate and 2-mercaptoethanol. The substrates azo-casein and the hide powder azure were used to determine the reduction of proteolytic activity of IC[ratio ]MC compared with the activity of crude extract and crude extract with iodoacetamide. The IC[ratio ]MC SDS-PAGE pattern shows a higher quantity of bands than extract with the inhibitor iodoacetamide or than crude extract. In addition, anti-<jats:italic>Entamoeba histolytica<\/jats:italic>antibodies from amoebic liver abscess patients recognized a richer antigenic Western blot pattern in the IC[ratio ]MC fraction than in crude extract alone or with inhibitor. The described method has proved to be suitable to preserve amoebic antigens for its use in diagnostic tests and it can be used for immunological response studies against<jats:italic>E. histolytica<\/jats:italic>antigens. Furthermore we propose that this method to obtain the IC[ratio ]MC fraction can be applied for the study of other microorganisms or cells with high enzymatic content.<\/jats:p>","DOI":"10.1017\/s0031182005007730","type":"journal-article","created":{"date-parts":[[2005,7,28]],"date-time":"2005-07-28T19:53:37Z","timestamp":1122580417000},"page":"231-236","source":"Crossref","is-referenced-by-count":8,"title":["Preparation of<i>Entamoeba histolytica<\/i>antigens without enzymatic inhibitors"],"prefix":"10.1017","volume":"131","author":[{"given":"M. S.","family":"FLORES","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"E.","family":"TAMEZ-TREVI\u00d1O","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"F.","family":"CASTA\u00d1EDA","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R.","family":"TIJERINA-MENCHACA","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"L.","family":"GALAN-WONG","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R.","family":"RANGEL","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"56","published-online":{"date-parts":[[2005,5,19]]},"reference":[{"key":"S0031182005007730_ref018","unstructured":"SA\u00cdD-FERN\u00c1NDEZ, S. & FLORES-CASTA\u00d1EDA, M. S. (1996).Diagn\u00f3stico molecular de amibiasis.Gaceta M\u00e9dica de M\u00e9xico 132,314\u2013318."},{"key":"S0031182005007730_ref016","doi-asserted-by":"publisher","DOI":"10.1017\/S0031182004005074"},{"key":"S0031182005007730_ref010","unstructured":"HARRIS, E. L. V. & ANGAL, S. (1995). Protein Purification Methods: A Practical Approach .IRL Press,Oxford."},{"key":"S0031182005007730_ref009","doi-asserted-by":"publisher","DOI":"10.1017\/S0031182004005086"},{"key":"S0031182005007730_ref007","unstructured":"FLORES-CASTA\u00d1EDA, M. S. (1999).Procedure to preserve antigens of Entamoeba histolytica without enzymatic inhibitors and their use in immunological methods. United States Patent 5 861 263."},{"key":"S0031182005007730_ref004","doi-asserted-by":"publisher","DOI":"10.1142\/9781848160583_0001"},{"key":"S0031182005007730_ref006","unstructured":"FLORES DE CASTA\u00d1EDA, M. S. (2002).Procedimiento para la preservaci\u00f3n de mol\u00e9culas antig\u00e9nicas, sin el uso de inhibidores enzim\u00e1ticos y su aplicaci\u00f3n en m\u00e9todos inmunol\u00f3gicos. Mexican Patent 209648."},{"key":"S0031182005007730_ref001","unstructured":"ARAIZA-OROZCO, L. M. , AVILA, E. , MUNOZ, M. L. & ARIAS-NEGRETE, S. (1999). Entamoeba histolytica: surface proteolytic activity and its relationship with in vitro virulence.Folia Parasitologica 46,161\u2013167."},{"key":"S0031182005007730_ref008","unstructured":"FLORES DE CASTA\u00d1EDA, M. S. (1995).Procedure to preserve antigens of E. histolytica without enzymatic inhibitors. United States Patent 5459042."},{"key":"S0031182005007730_ref005","unstructured":"FLORES DE CASTA\u00d1EDA, M. S. (2002).Procedimiento para la preservaci\u00f3n de mol\u00e9culas antig\u00e9nicas, sin el uso de inhibidores enzim\u00e1ticos. Mexican Patent 209646."},{"key":"S0031182005007730_ref012","doi-asserted-by":"publisher","DOI":"10.1139\/m77-062"},{"key":"S0031182005007730_ref015","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.13.2.196-206.2000"},{"key":"S0031182005007730_ref011","doi-asserted-by":"crossref","unstructured":"LOWRY, O. H. , ROSEBROUGH, N. J. , FARR, A. L. & RANDALL, R. J. (1951).Protein measurement with the Folin phenol reagent.Journal of Biological Chemistry 193,265\u2013275.","DOI":"10.1016\/S0021-9258(19)52451-6"},{"key":"S0031182005007730_ref020","doi-asserted-by":"crossref","unstructured":"TAMEZ-TREVI\u00d1O, E. , SEP\u00daLVEDA-SAAVEDRA, J. , TIJERINA-MENCHACA, R. & FLORES-CASTA\u00d1EDA, M. S. (2000).Systemic anti-amoeba antibodies induced orally in mice: detection by an ELISA test using preserved Entamoeba histolytica antigens without enzymatic inhibitors.Archives of Medical Research 31 (Suppl.),S28\u2013S29.","DOI":"10.1016\/S0188-4409(00)00130-2"},{"key":"S0031182005007730_ref019","doi-asserted-by":"publisher","DOI":"10.1006\/expr.1998.4258"},{"key":"S0031182005007730_ref014","doi-asserted-by":"publisher","DOI":"10.1016\/0166-6851(87)90133-2"},{"key":"S0031182005007730_ref013","doi-asserted-by":"publisher","DOI":"10.1126\/science.6162199"},{"key":"S0031182005007730_ref002","doi-asserted-by":"crossref","unstructured":"AVILA, E. , S\u00c1NCHEZ-GARZA, M. & CALDER\u00d3N, J. (1985). Entamoeba histolytica and E. invadens: Sulfhydril-dependent proteolytic activity.Journal of Protozoology 32,163\u2013166.","DOI":"10.1111\/j.1550-7408.1985.tb03032.x"},{"key":"S0031182005007730_ref003","unstructured":"CANTERA, C. S. , GOYA, L. , GARCIA, R. & SOFIA, A. (1999).Proteolytic activity assessment: use of the chromogenic substrate hide powder azure.Journal of the Society of Leather Technologists and Chemists 83,101\u2013106."},{"key":"S0031182005007730_ref017","doi-asserted-by":"publisher","DOI":"10.1016\/0009-8981(68)90127-7"}],"container-title":["Parasitology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.cambridge.org\/core\/services\/aop-cambridge-core\/content\/view\/S0031182005007730","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,7,14]],"date-time":"2021-07-14T04:06:47Z","timestamp":1626235607000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.cambridge.org\/core\/product\/identifier\/S0031182005007730\/type\/journal_article"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2005,5,19]]},"references-count":20,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2005,8]]}},"alternative-id":["S0031182005007730"],"URL":"https:\/\/doi.org\/10.1017\/s0031182005007730","relation":{},"ISSN":["0031-1820","1469-8161"],"issn-type":[{"value":"0031-1820","type":"print"},{"value":"1469-8161","type":"electronic"}],"subject":[],"published":{"date-parts":[[2005,5,19]]}}}